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DOI 10.1007/s11130-013-0360-2
ORIGINAL PAPER
Small differences in anthocyanin and volatile compounds content could be detected, and it may be of interest for the food
industry in order to classify a new individual into one of several
groups using different variables at once.
Keywords Roselle . Anthocyanins . Volatile compounds .
Categorical multifactorial analysis . Principal component
analysis (PCA) and factorial discriminate analysis (DFA)
Introduction
Hibiscus sabdariffa L. (Malvaceae) a plant native to Africa is
grown in tropical areas such as Sudan, Taiwan, Thailand and
Mexico [13]. It is a short-day annual shrub that is known by
different synonyms and common names such as roselle in
the U.S. and England, loiselle in France, jamaica or flor
de jamaica in Mexico and Spain, karkade in Sudan and
Arabia [4]. This plant grows over 180 cm; with globular
stems, leaves are ovate, flowers are grouped in terminal clusters, and petals are white with a reddish center at the base of
the staminal column. The calyx grows during maturity, fruit is
fleshy bright red, and the red persistent of calyx is the main
component having a sour taste, commonly used in preparation
of drinks, and as a food coloring [1].
In Mexico, it has recently gained an important position in
the market for soft drinks and commercial preparations.
Currently extracts of H. sabdariffa are marketed as supplements, because of the perception of their potential health
benefits [5, 6]. In traditional medicine it is used to treat kidney
stones [7], as a diuretic and has been studied extensively as an
antispasmodic [8], anti-inflammatory [9], treatment of atherosclerosis [10], antioxidant and anticancer treatment, bactericide and fungicide [5].
230
the TAC was calculated using Eq. 2, and the results were
expressed as mg of cyanidin 3-glucoside/L.
TAC
AB
D
MV V 100
eL
G
Where AB is absorbance; e is the molar extinction coefficient (molar absorptivity) of cyanidin 3-glucoside (26,900)
[18]; L is the cell length (1 cm); MW is anthocyanins
molecular weight (449.2); D is dilution factor; V is final volume
(mL); and G is the dry weight (dw) of roselle flour (mg).
Solid Phase Micro-Extraction (SPME)
For extraction of volatile compounds, ethanolic extracts of
roselle varieties were evaporated to dryness under an inert
nitrogen atmosphere.
A mixture of 1 g of sample and 10 ml of distilled water was
prepared in a 20 ml vial with 5 ml of a saturated NaCl solution
was added [16]. The vial was sealed airtight and incubated at
40 C for 20 min. Carboxen/PDMS of 65 m fiber (Supelco,
Bellefonte, PA) was used for the extraction of volatile compounds, in which the fiber needle was inserted and exposed in
the headspace of the vial for 20 min. Desorption was finally
performed by exposing the fiber for 4 min in the injection port
of the gas chromatograph.
Determination of Volatile Compounds by Gas
Chromatography (GC)
Gas chromatograph Varian 3800 GC (USA) was used and
analysis of volatile compounds was done according to the
methodology proposed by Pino et al. [13]. Separations were
performed using a CP-Wax 52 CB fused silica capillary
column, 30 m length and 0.25 m diameter. The initial
temperature was 60 C (2 min), then subsequently
programmed at 4 C/min to 250 C. Nitrogen was used as
carrier gas at a flow rate of 1 ml/min. Desoprtion temperature and detector was 250 C and injection was performed in
split mode (1:10). For identification of volatile compounds,
ethanol, geraniol, isoamylacetate, linalool, 2-nonanol, benzaldehyde, menthol, ethyl methyl phenylglycidate, and gamma undecalactone were used as reference standards, 2nonanol was used by the internal standard method.
Statistical Analysis
A categorical multifactorial experimental design with two
factors was used: variety (Negra, Sudan, Rosa and Blanca)
and type of extract (aqueous and ethanolic), with the following
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232
Conclusions
This study demonstrated evidence of differences between
anthocyanin and volatile compound contents of four varieties
with different degrees of pigmentation. Ethanol extracted
higher anthocyanins and volatile compounds, showing that
the major volatile compound found in both extracts was
geraniol. PCA and FDA allowed description of about 77 %
of variance values, allowing a clear classification of roselle
varieties using the first three components in a third dimension
scater plott. This model was supported by FDA correlation
confusion matrix. This could be useful for varieties classification according to their anthocyanin and volatile content and,
application of these techniques for natural and additive free
food industry is recommended.
Acknowledgments The authors thank to Colegio Superior de
Agricultura del Estado de Guerrero (CSAEGRO) and Quintn Obispo
Gonzlez for biological samples. One of the authors (GACM) also
acknowledges the scholarship from CONACYT-Mexico and SIP-IPN.
Conflict of interest The authors declare that they have no conflict of
interest.
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